6-amino-9-(substituted benzyl) purines and their n{11 oxides

ABSTRACT

Novel 6-amino-9-(substituted benzyl) purines are prepared by allowing an appropriate substituted benzyl halide, e.g., a substituted benzyl chloride, to react with adenine under the appropriate conditions. Oxidation of the resulting purine derivatives yields the corresponding N1-oxides. These compounds, i.e., both the purine derivatives and the N1-oxides thereof, have anti-coccidial activity and are useful for controlling cecal and/or intestinal coccidiosis when administered in minor quantities to animals, in particular to poultry, usually in admixture with animal sustenance.

United States Patent [1 1 Lira et al.

[ Nov. 5, 1974 I 6-AMINO-9-(SUBSTITUTED BENZYL) PURINES AND THEIR NOXIDES [75] Inventors: Emil P. Lira, Des Plaines; Walter M. Barker, Mundelcin; Robert C.

McCrae, Lake Zurich, all of III.

[73] Assignce: International Minerals & Chemical Corporation, Libcrtyville, Ill.

22 Filed: Feb. 4, 1972 21 Appl. No.: 219,848

Related US. Application Data [63] Continuation-in-part of Ser. No. 120,781, March 3,

1971, abandoned.

[52] US. Cl. 260/252, 424/253 [51] Int. Cl C07d 57/64 [58] Field of Search 260/252 [56] References Cited UNITED STATES PATENTS 3,213,095 l0/l965 Bambury et al. 260/252 FOREIGN PATENTS OR APPLICATIONS 45-27823 9/1970 Japan 260/252 OTHER PUBLICATIONS Schaeffer et al., J. of Medicinal Chemistry, Vol. 9,

Beilstein, Vol. 26, 1918, (p. 425).

Chemical Abstracts, Vol. 69, I968, (870]9j).

Montgomery et al., J. of Heterocyclic Chemistry, Vol.

I, No. 3. 1964, (p. [IS-I20).

Primary Examiner-Donald G. Daus Assistant Examiner-Anne Marie T. Tighe Attorney, Agent, or Firm-Peter Andress; James E. Wolber [57] ABSTRACT 27 Claims, No Drawings 6-AMINO-9-(SUBSTITUTED BENZYL) PURINES AND THEIR N OXIDES CROSS-REFERENCE TO RELATED APPLICATION BACKGROUND OF THE INVENTION This invention relates to new chemical compounds and the method of the preparation of the same. It relates further to the use of new compounds for treating and preventing coccidiosis. This invention still more particularly relates to novel 6-amino9-(substituted benzyl)purines and their corresponding N -oxides, and the use of the same in the control and treatment of coccidiosis.

Coccidiosis is a widespread poultry disease which is produced by infections of protozoa of the genus Eimeria which causes severe pathology in the intestines and ceca of poultry. Some of the most significant of these species are E. tenella, E. acervulina, E. necatrix, E. brunetti and E. maxima. This disease is generally spread by the birds picking up the infectious organism in droppings on contaminated litter or ground, or. by way of food drinking water. The disease is manifested by hemorrhage, accumulation of blood in the ceca, passage of blood in the droppings, weakness and digestive disturbances. The disease often terminates in the death of the animal, but the fowlwhich survive severe infections have had their market value substantially reduced as a result of the infection. Coccidiosis is, therefore, a

disease of great economic importance and extensive work has been done to find new and improved methods for controlling and treating coccidial infections in poultry.

SUMMARY OF THE INVENTION This invention is based on the discovery that certain novel 6-amino-9-(substituted benzyl)purines, as well as their corresponding N -oxides, have a surprisingly and unexpectedly high degree of activity against coccidiosis of poultry. Administering a small amount of at least one of these compounds, preferably in combination with I poultry feed, is effective to prevent or greatly reduce the incidence of coccidiosis. The compounds are effective against both the cecal form (caused by E. tenella) and the intestinal forms (principally caused by E. acervulina, E. brunetti, E. maxima and E. necatrix). The coccidiostats of this invention are particularly effective against the species that cause intestinal damage. In addition to preventing the pathology caused by coccidia, these compounds also exert an inhibitory effect on the oocysts by greatly reducing the number and/or the sporulation of those produced.

The novel purine derivatives of this invention are prepared by reacting an appropriate substituted benzyl halide and adenine in the presence of a base in a suitable reaction medium to obtain a novel 6-amino-9- (substituted benzyl)purine along with other isomers. The 9-isomer thus formed, either isolated or in the mixture with the other isomers, is then subjected to a suitable oxidizing agent such as hydrogen. peroxide, or a peraromatic or peraliphatic acid, to produce the novel corresponding 6-amino-9-(substituted benzyl)purine- N-oxide of this invention.

It is, therefore, a primary object of this invention to provide novel 6-amino-9-(substituted benzyl)purines and their corresponding N -oxides which are useful in the control of coccidiosis.

Another object of this invention is to provide novel anti-coccidial agents.

Still another object of this invention is to provide novel feed compositions useful for the prevention and suppression of coccidiosis in poultry.

A further object of this invention is to provide a new and useful method for the control of coccidiosis in poultry which comprises administering to the poultry minor amounts of the anti-coccidial substances of this invention.

A still further object of this invention is to provide a method for preparing novel 6-amino-9-(substituted benzyl)-purines and their corresponding N-oxides.

These and further objects of this invention will become apparent or be described as the description thereof herein proceeds.

DESCRIPTION OF THE PREFERRED EMBODIMENTS wherein n is 0 or 1, and (a) R and R independently are of the group consisting of hydrogen, halogen, i.e., chlorine, bromine, iodine or fluorine, nitro, and trihalomethyl, provided that at least one of such substituents is other than hydrogen and no more than one of such substituents is of the group consisting of nitro and trihalomethyl, and R and R are located in the 2 and 6 positions on the carbocyclic portion of the compound, or (b) R and R are both methyl and are located in the 3 and 4 positions on the carbocyclic ring. Preferred compounds are those where one of the substituents R and R is hydrogen or where both of the substituents are halogen, either the same or different, or methyl. Specific examples of novel compounds represented by the foregoing structural formula are 6-amino-9-(2- chlorobenzyl)purine; 6-amino- 9-( 3,4-dimethylbenzyl)purine; 6-amino-9-( 2- nitrobenzyl)purine; 6-amino-9-( 2- bromobenzyl)purine; 6 -amino-9-(2,6-dibromobenzyl) purine; 6-amino-9-(2-iodobenzyl)purine; 6-amino-9- (2-trifluoromethylbenzyl)purine; 6-amino-9-( 2-chloro- 6 -iodobenzyl)purine and 6-amino-9-(2-ehloro-6- bromobenzyl)purine; and their corresponding N- oxides. The most preferred compounds are 6-amino-9- (2,6-dichlorobenzyl)purine and 6-amino-9-(2-chloro- 6-fluorobenzyl)purine and their corresponding N- oxides, 6-amino- 9-(2,6-dichlorobenzyl)purine-N -oxide and 6-amino-9- 3 (2-chloro-6-fluorobenzyl )purine-N -oxide, tively.

In preparing the novel coccidiostats of this invention, the appropriate substituted benzyl halide (i.e., chloride, bromide, iodide or fluoride) is initially contacted with adenine in a suitable reaction medium, and the resulting reaction mixture is maintained to permit the reaction to take place. Useful benzyl halides may be characterized as those that are substituted so as to produce the novel compounds as hereinbefore described upon reaction with adenine, that is, benzyl halides containing one or two substituents of the group consisting of halogen, nitro and trihalomethyl in the 2 and 6 positions of the ring portion, provided no more than one of the substituents is of the group consisting of nitro and trihalomethyl, or containing methyl substituents in each of the 3 and 4 positions, with preferred benzyl halides being those containing only one of the aforementioned substituents in the 2-position (i.e., halogen, nitro and trihalomethyl) or containing two halogens or two methyl radicals. Benzyl halides that are useful in the preparation of the coccidiostats of this invention include 2,6- dichlorobenzyl chloride, 2-bromobenzyl bromide, 2- iodobenzyl bromide, 3,4-dimethylbenzyl chloride, 2- nitrobenzyl chloride, 2-chlorobenzyl chloride, and 2-chloro-6-fluorobenzyl bromide.

Suitable reaction media include both protic and aprotic solvents that are inert, i.e., non-reactive with the components of the reaction mixture under the reaction conditions that are maintained. Examples of such solvents include dialkyl alkanoamides, such as dimethyl formamide and dimethyl acetamide, alcohols, such as ethanol, propanol, isopropanol, butanol, amyl alcohol, cyclohexanol and benzyl alcohol, including aqueous systems containing up to about 50 percent of an alcohol such as previously characterized, dimethyl sulfoxrespecide, ethylene glycol and diethylene glycol. In general any of the solvents which are disclosed in the art as being useful in the base-catalyzed alkylation of adenine may be used in the first step of this invention for preparing the novel coccidiostats.

The bases that are useful for catalyzing the reaction between the substituted benzyl halide and adenine are those disclosed in the art as being useful for catalyzing the alkylation of adenine. In general, suitable bases include all those that have sufficient basicity to generate the adenine anion in the solvent system used. Examples of suitable bases include carbonates, e.g., alkali metal carbonates such as sodium carbonate and potassium carbonate, hydroxides, e.g., alkali metal .hydroxides such as sodium hydroxide, potassium hydroxide and lithium hydroxide, tertiary amines, e.g., trimethyl amine, triethyl amine, tripropyl amine and dimethyl analine, and alcoholates, e.g., potassium ethoxide, sodium ethoxide,potassium t-buto'xide and sodium ethylate. Preferred bases are those havinga pK, greater than 10.5 so that the adenine is substantially completely converted to its anion.

The relative proportions of the components of the reaction mixture of the first step may vary over a relatively wide range. The reactants may be used in stoichiometric amounts, i.e., equal moles of the two reactants may be used, or an excess, e.g., a percent or even greater molar excess, of either of the reactants may also be used. The quantity of the base is similarly not critical and may vary over a wide range, but it is preferably used in at least a stoichiometric amount with a slight excess being most preferred. The amount of the solvent used may also vary over a wide range. The solvent is used in a quantity sufficient to permit the reaction to proceed at a reasonable rate and facilitate isolation of the reaction product.

The reactants and the base catalyst are combined in the reaction medium in any conventional manner and in any order. As illustrative of a suitable manner of combining the components of the reaction mixture, the adenine is added to a solution of the base in the reaction medium, after which the substituted benzyl halide is then added, either neat or in a suitable solvent. Other methods of combining the reactants and catalyst will be obvious but it is preferred that they be combined so that the anion of the adenine forms no later than the time the benzyl halide is added and most preferably prior to such time, i.e., the benzyl halide is most preferably added last.

The reaction time and temperature conditions are not unduly critical. The time of the reaction will, however, decrease as the reaction temperature increases. The reaction will most conveniently be conducted between a temperature in the range of from about 50 to about C. However, substantially lower temperatures may be used so long as the reaction system is maintained in the liquid state. The reaction may be conducted at temperatures substantially higher than 100 C. provided the temperature is maintained at a level below that at which the components of the reaction mixture will decompose.

The reactants are thus contacted with each other in the presence of the base in the reaction medium to insure completion of the reaction. The product of the reaction, i.e., the 6-amino-9-(substituted benzyl)purine, either as a pure compound or along with the other isomers that form, e.g., the l-,3-, and 7-isomers, will either remain entirely in solution or precipitate after awhile, depending upon the quantity of solvent utilized, as hereinbefore outlined. Upon completion of the reaction, the reaction mixture may be cooled, e.g., to a temperature of about 0 C., to precipitate a solid product or to precipitate a further quantity of this product. The product is then isolated in the usual manner, such as by filtration, and, if desired, purified by conventional methods suchas washing with ethanol or water and recrystallizing from a suitable solvent such as acetic acid, aqueous acetic acid, dimethyl formamide or dimethyl sulfoxide. The 6-amino-9-(substituted benzyl)purine can be purified if it is used in that form as a coccidiostat in accordance with this invention instead of being first converted to the corresponding N-oxide.

The 6-amino-9-(substituted benzyl)purine thus obtained from the initial reaction may readily be converted to the corresponding novel purine-N -oxide of this invention by subjecting the same to the action of a suitable oxidizing agent. This is accomplished by initially dissolving the crude reaction product or isolated 9-isomer in a suitable acidic solvent, inorganic or organic, such a sulfuric acid, acetic acid, formic acid or butyric acid, dioxane, aqueous dioxane or ethylene glycol, and then adding an oxidizing agent that is compatible with the solvent. Oxidizing agents which may be used include hydrogen peroxide, peraromatic acids such as perbenzoic and metachloroperbenzoic acids, and peraliphatic acids such as peracetic and pertrifluoroacetic acids. The oxidizing mixture is either allowed to stand or is heated for a time sufficient to perdichlorobenzyl)purine mit the oxidizing reaction to substantially go to completion. Solid 6-amino-9-(substituted benzyl)purine-N- oxide, but not corresponding oxidized products of the other isomers, e.g., the 1, 3- and 7-isomers, is then recovered by cooling the oxidizing mixture to precipitate the product, or other suitable known methods of isolation may be used. Oxidized products of isomers other than the 9-isomers are not recovered, apparently since the other isomers are not oxidized or, if they are oxidized, the products thus formed are not easily isolated or they undergo further reactions to form other compounds that are not recovered with the oxidized 9- isomers. The crude product thus obtained is usually further purified by conventional methods such as recrystallization from an organic acid such as acetic acid.

The novel compounds of this invention are orally administered to poultry for the control and prevention of coccidiosis. Any number of conventional methods are suitable for administering the coccidiostats of this invention to poultry, as for example, they may be given in the poultry feed. The actual quantity of the coccidiostats administered to the poultry in accordance with this invention will vary over a wide range and be adjusted to individual needs, depending upon species of the coccidia involved and severity of the infection. The limiting criteria are that the minimum amount is sufficient to control coccidiosis and the maximum amount is such that the coccidiostat does not result in any undesirable effects.

A feed will typically contain from about 0.0005 to about 0.05 percent, preferably from about 0.0025 to about 0.01 percent, by weight of one of the coccidiostats of this invention. The optimum levels will naturally vary with the specific compound utilized and species of Eimeria involved, and can be readily determined by one skilled in the art. Levels of 6-amino-9-(2,6- dichlorobenzyl)purine and its corresponding N-oxide, which are among the most preferred coccidiostats of this invention, in poultry feed of from about 0.0035 percent to about 0.0075 percent by weight of the diet are especially useful in controlling the pathology associated with E. tenella, while the preferred concentration for similar control of intestinal-dwelling species is from about 0.0025 percent to about 0.0065 percent by weight of the diet. Amounts of about 0.0045 percent and 0.0065 percent by weight for 6-amino-9-(2,6- and 6-amino-9-(2,6- dichlorobenzyl)purine-N -oxide, respectively, are advantageous in reducing the pathogenic effects of both cecal and intestinal coccidiosis.

Depending on the compound employed, levels of 0.001 percent to 0.0035 percent possess the novel effects of reducing the number of oocysts passed in the droppings of infected chickens and/or inhibiting the subsequent division and maturation to infectivity, scientifically designated as the process of sporulation.

Thus, the combination of prevention of pathology, coupled with the inhibiting effect on the reproductive product of these organisms, the oocysts, present a unique two-fold method for the control of coccidiosis in poultry.

The quantity or concentration of a novel coccidiostat of this invention in any admixture in which it is administered to the poultry will, of course, vary in accordance with the type of admixture utilized.

Of the various methods of administering the coccidiostats of this invention to poultry, they are most conveniently administered as a component of a feed composition. The novel coccidiostats may be readily dispersed by mechanically mixing the same in finely ground form with the poultry feedstuff, or with an intermediate formulation (premix) that is subsequently blended with other components to prepare the final poultry feedstuff that is fed to the poultry. Typical components of poultry feedstuffs include molasses, fermentation residues, corn meal, ground and rolled oats, wheat shorts and middlings, alfalfa, clover and meat scraps, together with mineral supplements such as bone meal and calcium carbonate and vitamins.

The following non-limiting examples will serve to further illustrate the instant invention.

EXAMPLE l The compound 6-amino-9-(2,6- dichlorobenzyl)purine was initially prepared by slowly adding about 67.5 grams (0.5 mole) of dry adenine to a solution of about 57 grams (0.5 mole) potassium tbutoxide in one liter of absolute ethanol. This mixture was stirred for about two hours, and then about 97.5 grams (0.5 mole) of 2,6-dichlorobenzyl chloride was added over a period of about 4 hours. The resulting mixture was heated to reflux with stirring and allowed to react for about 42 hours, after which it was cooled to about lOC. The crude 6-amino-9- (2,6-dichlorobenzyl)purine thus formed was isolated by filtration and then washed with ice cold ethanol and ice cold water to yield 118 grams of a crude product.

The crude 6-amino-9-(2,6-dichlorobenzyl)purine obtained from the initial reaction was dissolved in one liter of acetic acid. A 30 percent aqueous solution-of hydrogen peroxide was then added in the amount of 350 milliliters. The solution was allowed to stand for 5 days at room temperature, after which about 400 milliliters of water was added and the solution was cooled. The resulting precipitate was recovered by filtration, reslurried in 500 milliliters of a 30 percent aqueous solution of acetic acid, and then in one liter of water. The solids recovered from the aqueous slurry were vacuum dried to yield grams of a crude 6-amino-9- (2,6-dichlorobenzyl)purine-N -oxide having a melting point of 29630l C., representing a 40 percent yield. An analytical sample of the same having a melting point of 298-302 C. was prepared by recrystallization from acetic acid.

EXAMPLE ll White Rock cockerels obtained from a commercial hatchery as day-old chicks were raised in a manner to prevent exposure to extraneous coccldial infection until they were fourteen days old. When the chicks were fourteen days old, they were individually weighed and divided into groups of ten each so that each group was substantially uniform in chick size and mean weight. The test compound, 6-amino-9-(2,6- dichlorobenzyl)purine-N -oxide prepared as described in Example I, was thoroughly dispersed in separate quantities of a vitamin K-deficient mash diet in concentrations of 0.01, 0.0075, 0.005, 0.0025 and 0.001 percent by weight. Each of these rations was offered ad libitium to a group of chicks. Other groups of chicks were fed the same mash diet without the addition of the 7 8 test compound. Two days after this diet was started, all TABLE II but three groups (30 chicks), designated the uninfected or normal control (UlC), were inoculated orally Medication N of wt. Gain Fecal Oocyst into the crop with 100,000 sporulatedoocysts of E. tenby ch'cks UIC 88%; ella. Two of the groups (20 chicks) which were so inoc- 5 None, UIC 30 100 159.8 0 0 ulated were fed the mash diet without any test com None [C 20 62,6 100 2.5 l 415 pound so as to serve as the mfected or positive control 0,00 10 559 g9 4 2,7 14

0.0025 10 71.6 114.4 1.7 170 (1C). The experiment was termlnated SIX days after the 0-005 10 93.8 1500 0 L22 chlcks were moculated. [0 0.0075 10 39.6 1431 0 0 Mortalities and fecal scores were recorded through- 12442 0 013 out the experiment. The fecal score was a grading on the extent of blood passed with droppings, using the following rating scale: EXAMPLE IV 0 i None 15 The procedure used in Example 11 was again followed 1 less than 10 percent except that the chicks were infected with five species 2 10 to 50 Percent of Eimeria simultaneously. Each chick was inoculated 3 Over 50 W with a total of 510,000 sporulated oocysts consisting of Oocysts counts were also obtained during the last two 10,000 E, ten lla, 200,000 E. acervulina, 50,000 E. days of the tests by microscopic examination of the necatrix, 100,000 E. brunetti, and 150,000 E. maxima. droppings to determine the average number of oocysts Table Ill which follows outlines the, results of this expassed per chick. At the end of the test, weight gains periment.

TABLE III Medication Oocyst by wt. No. of wt. Gain Hema- Mor- Fecal Count of Diet Chicks To UlC To [C tocrit tality Score (XIOS) None, 01c 100 458.7 31 0 0 0 None, 1c 20 21.8 100 24 3.0 24.30 0.001 10 29.8 137.0 20 3.0 32.55 0.0025 10 73.9 33901 27 40 2.7 47.02 0.005 10 93.1: 430.4 29 0 0.7 3.18 0.0075 10 92.9 426.1 32 0 0 0.24 001 10 83.4 382.6 31 0 0.3 0

and micro-hematocrits were determined. In determin- EXAMPLE V ing the mlcro-hematqcrlts, blood m d The general procedure outlined in Example II was lected from h chlck Wlth, a l p f again followed in this experiment, which was concapillary tube Whlch was then centnfugw to determme 40 ducted to compare the effectiveness of the test comthe percent by volume of packed blood cells. pound, i.e., .6-amino-9-(2,6-dichlorobenzyl)purine-N The results of this experiment are given in Table I. oxide, with five commercially available coccidiostats.

TABLE 1 Medication Oocyst by wt. No. of wt. Gain Hema- Mor- Fecal Count of Diet Chicks To UlC To lC tocrit tality Score (Xl0) None, UlC 30 100 181.9 31 0 0 0 None. lC 20 55.0 100 30 70 3.0 25.64 0.001 10 85.3 155.2 27 70 2.7 42.32 0.0025 10 37.9 69.0 27 40 2.3 56.95 0.005 10 86.3 156.9 28 0 2.3 31.0 0.0075 10 100.0 l8l.9 30 0 0.7 4.96 0.01 10 73.5 133.6 34 0 0 0.11

EXAMPLE Ill The test compound of this invention was administered This experiment was conducted substantially as outz s g l gfif gtz 'g g g ggzgg x ig g g gx lined in Example ll except that the chicks were inocualong with y concentratiopns utilized lated with 3,000,000 sporulated oocysts of E. acer- 60 vulina. The observations were limited to weight gains,

by wt. fecal scores and oocysts counts. Fecal scores were Com in Diet Chemical Name based on the amount of mucus and conslstency instead pound of on the blood in droppings, using the rating scale out- A 0.0125 l-(4-amino-2-N- r0 l-5- rinidi ll1nelcll 1n Exlample lIII. r d T bl n1 th l i-zioo1niiun i oh1o iidon i i n- T e resu ts o t is experiment are out me 1n a e e mi e 4 B 0.012s & Com oundA 1 M th 1-4- t 1 u as follows: 0.0004 ethox ybenzoat: us a y ace e 2 -Continued The severity of the infections was evaluated, again utilizing an uninfected or normal control (UIC) and an inby wt. fected or positive control (IC). There were 20 chicks Coin;1 in Diet Chem1cal Nam in both of these control groups and ten in each of the 5 remaining groups to which the test coccidiostats were c 0.00825 Ethyl-4-hydroxy-6,7-diisobutoxy-3- administered. The criteria used in evaluating the effec- D 00125 g g afgg gl h '4 l tiveness of the six anti-coccidial preparations being E @0125 j f g$ f gmg tested include weight gains along with at least one of the following observations: percent mortality, percent hematocrit and/or fecal score, as outlined in Example The compounds were administered to the chicks in feed for a total of nine days, which was the ext t of The results of this experiment are presented in Table this experiment; The chicks were inoculated, as in the IV as fOHOWSI TABLE IV None Test Commercial Coccidiostats Compound A B C D E E. tenella wt. gain to UlC 100.0 49.3 95.2 99.3 96.6 97.3 97.3 96.6 wt. ain to 1c 202.8 100.0 193.1 201.4 195.8 197.2 197.2 195.8 Mortality 0 55.0 0 0 0 0 0 0 Hematocrit 3O 28 28 28 30 30 E. acervulina wt. ain to UlC 100.0 45.3 100.0 89.0 98.3 100.0 86.6 54.6 wt. ain to 1c 220.5 100.0 220.5 196.2 216.7 220.5 191.0 120.5 Fecal Score 0 3 0 3 0 1 1 3 E necatrix wt. gain to UlC 100.0 39.2 95.6 56.0 72.8 70.2 93.7 57.0 wt. ain to 1c 254.8 100.0 243.5 141.9 185.5 179.0 238.7 145.2 Mortality 0 8O 0 0 0 0 0 0 E. brurlelti 7'8 w"t. ain't'o UIC 100.0 34.3 87.8 40.1 39.5 92.5 96.5 61.0 wt. ain to 1C 291.5 100.0 255.9 116.9 115.2 274.6' 281.4 178.0 Fecal Score 0 3 0 3 3 0 0 2 E. maxima wt. ain to UlC 100.0 60.0 89.5 73.3 73.8 86.7 74.8 64.3 wt. ain to 1c 166.7 100.0 149.2 122.2 123.0 144.4 124.6 107.1 Fecal Score 7 0 3 0 3 3 1 l 3 Mixed lnfect. 5 s "70 wt. gain to u1c 100.0 44.7 91.7 a 67.5 67.1 83.3 89.5 69.7 wt. ain to 1C 223.5 100.0 204.9 151.0 150.0 186.3 200.0 155.9 Mortality 0 40 0 0 0 0 0 0 Fecal Score 0 3 0 2 2 l l 3 previous examples, two days after the experiment EXAMPLE VI l of h compounds were evaluated 3 1 40 The procedures were essentially the same as in Exmdlvldu? Infections of each f 1 9 of Elmena amples II and 111, except the E. tenella trial was termiand a mufture 9? the five Specles glven Simultaneously nated five days post inoculation and the infected chicks The species utilized and number of sporulated oocysts were each given 200,000 sporulated oocysts of 5 mm admmlstered are hsted below' ella or 5,300,000 sporulated oocysts of E. acervulina.

individual The results, shown in Table V demonstrate the anticocgifect ion f gt ixg ln f p cidial activity of the test compound, 6-amino-9-(2,6- pews Pwes dichlorobenzyl)purine, as compared with the commer- E. mien" 75.000 10,000 cially available coccidiostats which are identified as f iz fg' 383 888 8 888 compounds B (l-(4-am1no-2-N-propyl-5-pyrinidinyl- ,1 zoojooo 1001000 methyl)-2-picolinium chloride'hydrochloride plus m xim 2 T t I ..i%.%8 methyl-4-acetamide-2-ethoxybenzoate) and D (3,5-

dichloro-2,6-dimethyl-4-pyridinol) in Example V.

TABLE V E. acervullna i Medlcatlon Totul Medication bWL No. of Aver. wt. Guln Fecal ln Diet of let Chicks Gain(g) 0 0 Score None. UlC 5 218 100 144.4 0 None. lC S 151 68.8 100 3 Compound B 0.0125 5 221 101.4 146.4 0 Test Compound 0.0035 5 223 102.3 147.7 0

E. tenella I Medication Medication No. of Total %Mor- Hemaln Diet by Wt. Chicks Aver. wt. Gain tocrit tality of Diet Gain(g) o o C None. UlC 5 198 100 176.8 33 0 None, lC 5 112 56.5 100 14 Compound 0.0125 5 203 102.5 181.2 33 0 D Test Compound 0.0045 10 197 99.5 175.9 32 0 EXAMPLE VII The procedures employed in Example VI were again followed except that 125,000 sporulated oocysts of E. tenella were given to the infected chicks. The E. acervulina infection was the same. The efficacy of 6-amino- 9-(2-chloro-6-fluorobenzyl)purine-N -oxide (test compound) is compared to that of compound B of Example I V against both species. The results are given in Table VI.

TABLE VI E. Ienella Medication Total Medication by Wt. No. of Aver. wt. Gain H %Moremaln Diet of Diet Chicks Gain(g) to UIC To IC tocrit tality None, UlC 5 189 100 148.8 32 None, 1C 127 67.2 100 16 50 Compound 0.0125 5 186 99.6 146.6 0 B Test 0.005 5 203 107.4 159.8 30 0 Compound E. acervulina Medication Total Medication by Wt. No. of Aver. wt. Gain Fecal In Diet of Diet Chicks Gain(g) To UlC To 1C Score None, UlC 5 224 100 138.3 0 None, IC 5 162 72.3 100 3 Compound B 0.0125 5 218 97.3 134.6 1

Test Compound EXAMPLE VIII In order to demonstrate the effect of 6-amino-9-(2,6- dichlorobenzyl)purine (Test Compound A) and its corresponding N -oxide (Test Compound B) on oocysts the following procedures were utilized.

Two groups of five chicks (same type as used in Example II) were each assigned to each of 14 treatments. Feed containing various levels of either of Test Compounds A or B was fed to the birds 24 hours prior to their infection with E. acervulina. Unrnedicated feed 30 dichromate and homogenized from which a 200 gram TABLE VIl Aver. No. Oocyst of Total Medication of Oocyst Produc- Sample Spor- Reduction of and 1n X l0lchick tion To Sporuulation to Sporulated The Diet /day Controls lated Controls Oocysts Test Compound A None 35.728 100.00 36.71 100.00 0.0045 0.051 0.14 0 0 100.00 0.0035 1.200 3.36 0 0 100.00 0.0025 9.589 26.84 2.90 7.90 97.88 0.0015 16.858 47.84 5.36 11.24 94.62 0.001 14.151 39.61 6 67 16.84 93.33 gest Compound None 109.77 100.00 22.5 100.00 0.0065 0.33 0.30 0 0 100.00 0.0055 0.88 0.80 O 0 100.00 0.0045 0.39 0.36 0 0 100.00 0.0035 10.70 9.75 0.6 2.67 99.74 0.0025 168.89 153.82 2.1 9.33 86.65 0.0015 77.35 71.38 3.1 13.78 90.16 0.001 107.35 97.81 6.0 26.67 73.91

minus Z oocyst production to controls X k sporulution to controls total 7: reduction of sporuluted oocysts.

High number possibly due to sumpling or counting error.

was given to four groups of control chicks for the same period. The procedures employed with both test compounds were identical except for the number of days It will be apparent to one skilled in the art from the foregoing that the novel 6-amino-9-(substituted benzyl)purine s and corresponding N -oxides of this invention are effective coccidiostats. This is especially apparent from the results of experiments outlined in the foregoing examples. In these tests, the preferred coccidiostat of this invention, i.e., 6-amino-9- (2,6-dichlorobenzyl)purine and its corresponding N- oxide, administered to the test chicks in a mash diet was effective in controlling coccidiosis when the chicks were inoculated with sporulated oocysts of species of Eimeria. The test compound was effective against both the cecal form and the intestinal form of coccidiosis. The test compound also compared very favorably with five compounds which are currently commercially available coccidiostats.

Although this invention has been described in relation to specific embodiments, it will be apparentthat obvious modifications may be made by one skilled in the art without departing from the intended scope thereof as defined by the appended claims.

We claim:

1. A compound of the structural formula:

wherein n is or 1, and one of R and R is hydrogen with the other being a halogen selected from the group consisting of chlorine, bromine and iodine.

2. A compound in accordance with claim 1 wherein n is O.

3. A compound in accordance with claim 1 wherein n is l.

4. A compound of the structural formula:

wherein n is 0 or 1 and both R and R are halogen.

5. A compound in accordance with claim 4 wherein n is O. i

6. A compound in accordance with claim 4 wherein n is l.

7. A compound in accordance with claim 5 wherein one of R and R is chlorine with the other being fluorine.

8. A compound in accordance with claim 6 wherein one of R and R is chlorine with the other being fluorine.

9. A compound in accordance with claim 5 wherein one of R and R is chlorine with the other being bromine.

10. A compound in accordance with claim 6 wherein one of R and R is chlorine with the other being bromine.

11. A compound in accordance with claim 5 wherein one of R and R is chlorine with the other being iodine.

12. A compound in accordance with claim 6 wherein one of R and R is chlorine with the other being iodine.

13. A compound of the structural formula:

NII:

6-amino-9-( 3 ,4-dimethylbenzyl )purine. 6-amino-9-(3,4-dimethylbenzyl)purine-N -oxide.

6-amino-9-(2-chlorobenzyl)purine. 6-amino-9-(2-chlorobenzyl)purine-N -oxide. 6-amino-9-(2-bromobenzyl)purine. 6-amino-9-(2-bromobenzyl)purine-N -oxide.

24. 6-amino-9-(2-iodobenzyl)purine.

25. 6-amino-9-(2-iodobenzyl)purine-N-oxide.

26. 6-amino-9-(2-trifluoromethylbenzyl)purine.

27. 6-amino-9-( Z-trifluoromethylbenzyl )purine-N oxide. 

1. A COMPOUND OF THE STRUCTURE FORMULA
 2. A compound in accordance with claim 1 wherein n is
 0. 3. A compound in accordance with claim 1 wherein n is
 1. 4. A compound of the structural formula:
 5. A compound in accordance with claim 4 wherein n is
 0. 6. A compound in accordance with claim 4 wherein n is
 1. 7. A compound in accordance with claim 5 wherein one of R1 and R2 is chlorine with the other being fluorine.
 8. A compound in accordance with claim 6 wherein one of R1 and R2 is chlorine with the other being fluorine.
 9. A compound in accordance with claim 5 wherein one of R1 and R2 is chlorine with the other being bromine.
 10. A compound in accordance with claim 6 wherein one of R1 and R2 is chlorine with the other being bromine.
 11. A compound in accordance with claim 5 wherein one of R1 and R2 is chlorine with the other being iodine.
 12. A compound in accordance with claim 6 wherein one of R1 and R2 is chlorine with the other being iodine.
 13. A compound of the structural formula:
 14. A compound in accordance with claim 13 wherein n is
 0. 15. A compound in accordance with claim 13 wherein n is
 1. 16. 6-amino-9-(2,6-dichlorobenzyl)purine.
 17. 6-amino-9-(2,6-dichlorobenzyl)purine-N1-oxide.
 18. 6-amino-9-(3,4-dimethylbenzyl)purine.
 19. 6-amino-9-(3,4-dimethylbenzyl)purine-N1-oxide.
 20. 6-amino-9-(2-chlorobenzyl)purine.
 21. 6-amino-9-(2-chlorobenzyl)purine-N1-oxide.
 22. 6-amino-9-(2-bromobenzyl)purine.
 23. 6-amino-9-(2-bromobenzyl)purine-N1-oxide.
 24. 6-amino-9-(2-iodobenzyl)purine.
 25. 6-amino-9-(2-iodobenzyl)purine-N1-oxide.
 26. 6-amino-9-(2-trifluoromethylbenzyl)purine.
 27. 6-amino-9-(2-trifluoromethylbenzyl)purine-N1-oxide. 